核磷蛋白A型突变体对髓性白血病细胞系增殖的影响  

Effect of type A mutant of nucleophosmin on proliferation of myelocytic leukemia cell lines

作　　者：骆展鹏[1] 何鹏[1] 张伶[1] 杨松[1] 钟晓明[1] 高玉洁[1] 

机构地区：[1]重庆医科大学医学检验系临床血液教研室,临床检验诊断学省部共建教育部重点实验室,重庆市重点实验室,重庆400016

出　　处：《临床检验杂志》2008年第1期20-23,共4页Chinese Journal of Clinical Laboratory Science

基　　金：重庆市教委科学技术研究基金资助项目(No.KJ050309)

摘　　要：目的研究核磷蛋白A型突变体基因转染对人髓性白血病细胞系体外增殖的影响。方法构建含核磷蛋白A型突变基因(NPM-mA)的真核表达质粒pEGFPC1-NPM-mA,选择NPM-mA阴性、抑癌基因p14ARF有无缺失的两株人髓性白血病细胞KG-1a和K562作为靶细胞,将pEGFPC1-NPM-mA重组质粒和pEGFPC1空质粒分别转染白血病细胞,以未转染组为对照。用RT-PCR和免疫组化来分析转染细胞目的基因mRNA和蛋白质的表达。采用台盼蓝拒染法计数活细胞,绘制转染前后生长曲线,观察细胞体外生长的改变。结果转染后的两种白血病细胞株表达NPM-mA mRNA,胞浆内NPM-mA蛋白阳性,符合NPM突变蛋白胞质移位的特点。细胞生长曲线显示,转染NPM-mA质粒的KG-1a细胞较空质粒转染和未转染组细胞生长速度明显加快,而转染NPM-mA质粒的K562细胞则较对照组细胞生长明显受抑制(P<0.05)。结论NPM-mA基因稳定转染可影响白血病细胞的增殖活性,在无ARF缺失时NPM-mA突变体可促进白血病细胞体外生长。Objective To investigate the effect of type A mutant of nucleophosmin （NPM-mA） on the proliferation of human myelocytic leukemia cell lines in vitro. Methods The eukaryotic expression plasmid pEGFPC1-NPM-mA containing NPM-mA gene was constructed. Two NPM-mA-negtive leukemic cell lines （KG-1a and K562） were chosen as research objects, which expressed anti-oncogene p14 alternative reading frame （ARF） or not respectively. After stably transfection of the two cells with pEGFPC1-NPM-mA and empty plasmid pEGFPC1, RT-PCR and immunohistochemistry were used to detect the expression of mRNA and protein of NPM-mA respectively. The untransfected group was served as control. The proliferation effect was assessed by viable cell count （trypan blue） and cell growth curve. Results The expression of NPM-mA mRNA was detectable in transfected cells. NPM-mA protein was located in cytoplasm. The growth curve showed that after transfection of pEGFPC1-NPM-mA, KG-1 a cells displayed high growth rate compared with empty plasmid transfected or untransfected group, however the growth of pEGFPC1-NPM-mA-transfected K562 cells were significantly suppressed compared with control groups （ P 〈 0.05 ）. Conclusion The expression of NPM-mA may have effect on the proliferation of leukemia cells, and even if absence of ARF NPM-mA can promote the growth of leukemic cells in vitro.

关 键 词：白血病 核磷蛋白 基因突变 

分 类 号：R733.7[医药卫生—肿瘤]